{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,21]],"date-time":"2025-10-21T00:47:54Z","timestamp":1761007674590,"version":"build-2065373602"},"reference-count":74,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2025,10,14]],"date-time":"2025-10-14T00:00:00Z","timestamp":1760400000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Hidalgo Fund for Science and Technology","award":["DDCTI\/FH\/DCI-097"],"award-info":[{"award-number":["DDCTI\/FH\/DCI-097"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Corn head smut is a disease caused by the fungus Sporisorium reilianum. Chemical treatments and tolerant hybrids are available for control of this disease; however, these can lead to the development of resistant strains, complicating its management. This microorganism produces two extracellular enzymes\u2014aspartyl protease EAP1 and xylanase SRXL1\u2014which may be involved in the host penetration and colonization processes. Plants produce peptides that inhibit enzymes involved in phytopathogenesis, which could serve as tools to control plant pathogens. In this study, enzyme inhibitors were extracted from corn seed flours derived from two hybrids\u2014a white variety (DK-2061) and a purple variety (BOGUI)\u2014with the objective of evaluating their inhibitory effects on the enzymes EAP1 and SRXL1. Interestingly, the identified inhibitors were starches that showed 100% enzymatic inhibition. These compounds were characterized through Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDS) analysis. The purified starches exhibited acetylation (1.52 \u00b1 0.07% for DK-2061 and 1.16 \u00b1 0.04% for BOGUI) as a result of the purification process, due to the use of an acetate regulator; however, they maintained their complete inhibitory activity against the studied enzymes. In contrast, the activity of the purified inhibitors was lost after incubation with \u03b1-amylase. Each isolated compound showed uncompetitive inhibition on both enzymatic activities, indicated by a decrease in Km and Vmax values, as determined using the Lineweaver-Burk equation. This represents the first report of the inhibitory effects of corn starches on aspartyl protease and xylanase extracted from S. reilianum. Therefore, these compounds could serve as valuable elements in strategies to manage head smut, potentially reducing the reliance on chemical fungicides.<\/jats:p>","DOI":"10.3390\/ijms26209974","type":"journal-article","created":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T15:20:12Z","timestamp":1760973612000},"page":"9974","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Enzymatic Inhibitors of Aspartyl Protease EAP1 and Xylanase SRXL1 from Sporisorium reilianum Isolated from Corn Seeds"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-5862-9564","authenticated-orcid":false,"given":"Yusiri","family":"Vel\u00e1zquez-Ju\u00e1rez","sequence":"first","affiliation":[{"name":"Laboratorio Nacional Conahcyt de Investigaci\u00f3n y Servicios para la Productividad del Campo, Universidad Polit\u00e9cnica de Pachuca, Carretera Pachuca-Cd. Sahag\u00fan km 20, Ex-Hacienda de Santa B\u00e1rbara, Zempoala 43830, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5491-3679","authenticated-orcid":false,"given":"Alejandro","family":"T\u00e9llez-Jurado","sequence":"additional","affiliation":[{"name":"Laboratorio Nacional Conahcyt de Investigaci\u00f3n y Servicios para la Productividad del Campo, Universidad Polit\u00e9cnica de Pachuca, Carretera Pachuca-Cd. Sahag\u00fan km 20, Ex-Hacienda de Santa B\u00e1rbara, Zempoala 43830, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8164-2835","authenticated-orcid":false,"given":"Macaria","family":"Hern\u00e1ndez-Ch\u00e1vez","sequence":"additional","affiliation":[{"name":"Laboratorio de Optomecatr\u00f3nica y Energ\u00edas, Unidad Profesional Interdisciplinaria de Ingenier\u00eda Campus Hidalgo (UPIIH), Instituto Polit\u00e9cnico Nacional, Distrito de Educaci\u00f3n, Salud, Ciencia, Tecnolog\u00eda e Innovaci\u00f3n, San Agust\u00edn Tlaxiaca 42162, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0055-2991","authenticated-orcid":false,"given":"Lourdes","family":"Villa-Tanaca","sequence":"additional","affiliation":[{"name":"Laboratorio de Biolog\u00eda Molecular de Bacterias y Levaduras, Departamento de Microbiolog\u00eda, Escuela Nacional de Ciencias Biol\u00f3gicas, Instituto Polit\u00e9cnico Nacional, Prol. de Carpio y Plan de Ayala, Col. Sto. Tom\u00e1s, Ciudad de M\u00e9xico 11340, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Martha Patricia","family":"Falc\u00f3n-Le\u00f3n","sequence":"additional","affiliation":[{"name":"Laboratorio Nacional Conahcyt de Investigaci\u00f3n y Servicios para la Productividad del Campo, Universidad Polit\u00e9cnica de Pachuca, Carretera Pachuca-Cd. Sahag\u00fan km 20, Ex-Hacienda de Santa B\u00e1rbara, Zempoala 43830, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3278-2783","authenticated-orcid":false,"given":"Yuridia","family":"Mercado-Flores","sequence":"additional","affiliation":[{"name":"Laboratorio Nacional Conahcyt de Investigaci\u00f3n y Servicios para la Productividad del Campo, Universidad Polit\u00e9cnica de Pachuca, Carretera Pachuca-Cd. Sahag\u00fan km 20, Ex-Hacienda de Santa B\u00e1rbara, Zempoala 43830, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Jiao, Y., Chen, H.D., Han, H., and Chang, Y. (2022). Development and utilization of corn processing by-products: A review. Foods, 11.","DOI":"10.3390\/foods11223709"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Mohammed, A.A.B.A., Hasan, Z., Omran, A.A.B., Kumar, V.V., Elfaghi, A.M., Ilyas, R.A., and Sapuan, S.M. (2022). Corn: Its structure, polymer, fiber, composite, properties, and applications. Polymers, 14.","DOI":"10.3390\/polym14204396"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1094\/Phyto-75-577","article-title":"Factors affecting the development of head smut caused by Sphacelotheca reiliana on corn","volume":"75","author":"Matyac","year":"1985","journal-title":"Phytopathology"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1146\/annurev-phyto-082718-100139","article-title":"Molecular interactions between smut fungi and their host plants","volume":"57","author":"Zuo","year":"2019","journal-title":"Annu. Rev. Phytopathol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1080\/15572536.2003.11833215","article-title":"The biological cycle of Sporisorium reilianum f. sp. zeae: An overview using microscopy","volume":"94","author":"Martinez","year":"2002","journal-title":"Mycologia"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1317","DOI":"10.1128\/EC.4.8.1317-1327.2005","article-title":"Mating type loci of Sporisorium reilianum: Novel pattern with three a and multiple b specificities","volume":"4","author":"Schirawski","year":"2005","journal-title":"Eukaryot. Cell"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2037","DOI":"10.1104\/pp.111.179499","article-title":"Sporisorium reilianum infection changes inflorescence and branching architectures of maize","volume":"156","author":"Ghareeb","year":"2011","journal-title":"Plant Physiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"77","DOI":"10.21273\/HORTSCI.19.1.77","article-title":"Effects of cultural practices on the incidence of head smut in sweet corn","volume":"19","author":"Mack","year":"1984","journal-title":"HortScience"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1080\/01140671.2006.9514383","article-title":"Fungicide control of head smut (Sporisorium reilianum) of sweetcorn (Zea mays)","volume":"34","author":"Wright","year":"2006","journal-title":"N. Z. J. Crop Hortic. Sci."},{"key":"ref_10","first-page":"e200","article-title":"Sensibilidad del carb\u00f3n de la espiga del ma\u00edz a diferentes fungicidas","volume":"30","author":"Salinas","year":"2023","journal-title":"CIENCIA Ergo-Sum"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Islam, T., Danishuddin Tamanna, N.T., Matin, M.N., Barai, H.R., and Haque, M.A. (2024). Resistance mechanisms of plant pathogenic fungi to fungicide, environmental impacts of fungicides, and sustainable solutions. Plants, 13.","DOI":"10.3390\/plants13192737"},{"key":"ref_12","first-page":"39","article-title":"Resistance of varieties and hybrids of corn (Zea mays) to Sporisorium reilianum and grain yield","volume":"29","year":"2011","journal-title":"Rev. Mex. Fitopatol."},{"key":"ref_13","first-page":"534","article-title":"Genetic resistance to Sporisorium reilianum f. sp. zeae in selected maize (Zea mays L.) lines with white and yellow endosperm","volume":"35","year":"2017","journal-title":"Rev. Mex. Fitopatol."},{"key":"ref_14","unstructured":"Sultan, S. (2016). Stenocarpella maydis and Sporisorium reilianum: Two pathogenic fungi of maize. Fungal Pathogenicity, INTECH."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.biocontrol.2014.04.011","article-title":"Application of Bacillus subtilis in the biological control of the phytopathogenic fungus Sporisorium reilianum","volume":"76","year":"2014","journal-title":"Biol. Control"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Gonz\u00e1lez-Le\u00f3n, Y., De la Vega-Camarillo, E., Ram\u00edrez-Vargas, R., Anducho-Reyes, M.A., and Mercado-Flores, Y. (2024). Whole genome analysis of Bacillus velezensis 160, biological control agent of corn head smut. Microbiol. Spectr., 12.","DOI":"10.1128\/spectrum.03264-23"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Zhang, S., Li, C., Si, J., Han, Z., and Chen, D. (2022). Action mechanisms of effectors in plant-pathogen interaction. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23126758"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1016\/j.pbi.2008.04.007","article-title":"Enzyme-inhibitor interactions at the plant-pathogen interface","volume":"11","year":"2008","journal-title":"Curr. Opin. Plant Biol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Rodr\u00edguez-Sifuentes, L., Marszalek, J.E., Chuck-Hern\u00e1ndez, C., and Serna-Sald\u00edvar, S.O. (2020). Legumes protease inhibitors as biopesticides and their defense mechanisms against biotic factors. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21093322"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Del Prete, S., and Pagano, M. (2024). Enzyme inhibitors as multifaceted tools in medicine and agriculture. Molecules, 29.","DOI":"10.3390\/molecules29184314"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1016\/j.bbrc.2005.09.030","article-title":"A family 11 xylanase from the pathogen Botrytis cinerea is inhibited by plant endoxylanase inhibitors XIP-I and TAXI-I","volume":"337","author":"Brutus","year":"2005","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1016\/j.bbrc.2004.12.036","article-title":"Cloning and characterization of two endoxylanases from the cereal phytopathogen Fusarium graminearum and their inhibition profile against endoxylanase inhibitors from wheat","volume":"327","author":"Volckaert","year":"2005","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1042\/BJ20061291","article-title":"TLXI, a novel type of xylanase inhibitor from wheat (Triticum aestivum) belonging to the thaumatin family","volume":"403","author":"Fierens","year":"2007","journal-title":"Biochem. J."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1464","DOI":"10.1094\/MPMI-04-13-0121-R","article-title":"Constitutive expression of the xylanase inhibitor TAXI-III delays Fusarium head blight symptoms in durum wheat transgenic plants","volume":"26","author":"Moscetti","year":"2013","journal-title":"MPMI"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"815","DOI":"10.1094\/MPMI-07-16-0148-R","article-title":"PvPGIP2 accumulation in specific floral tissues but not in the endosperm limits Fusarium graminearum infection in wheat","volume":"29","author":"Tundo","year":"2016","journal-title":"Mol. Plant Microbe Interact."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.femsle.2004.09.019","article-title":"An endo-polygalacturonase (PG) of Fusarium moniliforme escaping inhibition by plant polygalacturonase-inhibiting proteins (PGIPs) provides new insights into the PG-PGIP interaction","volume":"240","author":"Sella","year":"2004","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.pmpp.2005.10.002","article-title":"Polygalacturonase-inhibiting protein 2 of Phaseolus vulgaris inhibits BcPG1, a polygalacturonase of Botrytis cinerea important for pathogenicity, and protects transgenic plants from infection","volume":"67","author":"Manfredini","year":"2005","journal-title":"Physiol. Mol. Plant Pathol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1380","DOI":"10.1104\/pp.105.067546","article-title":"The polygalacturonase-inhibiting protein PGIP2 of Phaseolus vulgaris has evolved a mixed mode of inhibition of endopolygalacturonase PG1 of Botrytis cinerea","volume":"139","author":"Sicilia","year":"2005","journal-title":"Plant Physiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2916","DOI":"10.1016\/j.foodchem.2013.04.013","article-title":"A sorghum xylanase inhibitor-like protein with highly potent antifungal, antitumor and HIV-1 reverse transcriptase inhibitory activities","volume":"141","author":"Lin","year":"2013","journal-title":"Food Chem."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Kumar, S., Singh, N., Mishra, B., Dube, D., Sinha, M., Singh, S.B., Dey, S., Kaur, P., Sharma, S., and Singh, T.P. (2010). Modulation of inhibitory activity of xylanase-\u03b1-amylase inhibitor protein (XAIP): Binding studies and crystal structure determination of XAIP-II from Scadoxus multiflorus at 1.2 \u00c5 resolution. BMC Struct. Biol., 10.","DOI":"10.1186\/1472-6807-10-41"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1007\/s11240-014-0590-5","article-title":"Overexpression of the RIXI xylanase inhibitor improves disease resistance to the fungal pathogen, Magnaporthe oryzae, in rice","volume":"120","author":"Hou","year":"2015","journal-title":"PCTOC"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1877","DOI":"10.1016\/j.drudis.2019.05.026","article-title":"Peptide-based protease inhibitors from plants","volume":"24","author":"Hellinger","year":"2019","journal-title":"Drug Discov. Today"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Gutierrez-Gongora, D., and Geddes-McAlister, J. (2021). From naturally-sourced protease inhibitors to new treatments for fungal infections. J. Fungi, 7.","DOI":"10.3390\/jof7121016"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Parisi, M.G., Oz\u00f3n, B., Vera, G.S.M., Garc\u00eda-Pardo, J., and Obreg\u00f3n, W.D. (2024). Plant protease inhibitors as emerging antimicrobial peptide agents: A comprehensive review. Pharmaceutics, 16.","DOI":"10.3390\/pharmaceutics16050582"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"668","DOI":"10.1126\/science.101.2635.668","article-title":"Crystallization of a trypsin inhibitor from soybean","volume":"101","author":"Kunitz","year":"1945","journal-title":"Science"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"15593258221131462","DOI":"10.1177\/15593258221131462","article-title":"Trypsin inhibitor isolated from Glycine max (soya bean) extraction, purification, and characterization","volume":"20","author":"Khan","year":"2022","journal-title":"Dose-Response"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1046\/j.1432-1327.1998.2540160.x","article-title":"Purification, characterization and cloning of an aspartic proteinase inhibitor from squash phloem exudate","volume":"254","author":"Christeller","year":"1998","journal-title":"Eur. J. Biochem."},{"key":"ref_38","first-page":"585","article-title":"A Kunitz trypsin inhibitor from chickpea (Cicer arietinum L.) that exerts an antimicrobial effect on Fusarium oxysporum f.sp. ciceris","volume":"4","author":"Nair","year":"2013","journal-title":"Agric. Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1080\/10826068.2013.835733","article-title":"Isolation of a trypsin-chymotrypsin inhibitor and its functional properties","volume":"44","author":"Wang","year":"2014","journal-title":"Prep. Biochem. Biotechnol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1016\/j.bbrc.2006.05.186","article-title":"Purification and characterization of a heat-stable serine protease inhibitor from the tubers of new potato variety \u201cGolden Valley\u201d","volume":"346","author":"Kim","year":"2006","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1016\/j.pep.2013.10.003","article-title":"Biochemical study of the extracellular aspartyl protease Eap1 from the phytopathogen fungus Sporisorium reilianum","volume":"92","year":"2013","journal-title":"Protein Expr. Purif."},{"key":"ref_42","first-page":"5309","article-title":"Purification and characterization of xylanase SRXL1 from Sporisorium reilianum grown in submerged and solid-state fermentation","volume":"8","year":"2013","journal-title":"BioRes"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1111\/nph.15596","article-title":"Origin and evolution of the plant immune system","volume":"222","author":"Han","year":"2019","journal-title":"New Phytol."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Han, Z., and Schneiter, R. (2024). Dual functionality of pathogenesis-related proteins: Defensive role in plants versus immunosuppressive role in pathogens. Front. Plant Sci., 15.","DOI":"10.3389\/fpls.2024.1368467"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Delaunois, B., Jeandet, P., Cl\u00e9ment, C., Baillieul, F., Dorey, S., and Cordelier, S. (2014). Uncovering plant-pathogen crosstalk through apoplastic proteomic studies. Front. Plant Sci., 5.","DOI":"10.3389\/fpls.2014.00249"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"622","DOI":"10.1134\/S2079059716050130","article-title":"Plant cell wall and mechanisms of resistance to pathogens","volume":"6","author":"Smirnova","year":"2016","journal-title":"Russ. J. Genet. Appl. Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1080\/07352689.2020.1757829","article-title":"The lifecycle of the plant immune system","volume":"39","author":"Li","year":"2020","journal-title":"Crit. Rev. Plant Sci."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"2008","DOI":"10.1128\/AAC.45.7.2008-2017.2001","article-title":"Novel bifunctional inhibitor of xylanase and aspartic protease: Implications for inhibition of fungal growth","volume":"45","author":"Dash","year":"2001","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Tundo, S., Mandal\u00e0, G., Sella, L., Favaron, F., Bedre, R., and Kalunke, R.M. (2022). Xylanase inhibitors: Defense players in plant immunity with implications in agro-industrial processing. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232314994"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Cotabarren, J., Lufrano, D., Parisi, M.G., and Obreg\u00f3n, W.D. (2020). Biotechnological, biomedical, and agronomical applications of plant protease inhibitors with high stability: A systematic review. Int. J. Exp. Plant Biol., 292.","DOI":"10.1016\/j.plantsci.2019.110398"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"560","DOI":"10.1104\/pp.103.024810","article-title":"Molecular cloning and functional analysis of a novel type of Bowman-Birk inhibitor gene family in rice","volume":"133","author":"Qu","year":"2003","journal-title":"Plant Physiol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1016\/j.phytochem.2005.11.010","article-title":"A Kunitz proteinase inhibitor from Archidendron ellipticum seeds: Purification, characterization, and kinetic properties","volume":"67","author":"Bhattacharyya","year":"2006","journal-title":"Phytochemistry"},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Tundo, S., Paccanaro, M.C., Elmaghraby, I., Moscetti, I., D\u2019Ovidio, R., Favaron, F., and Sella, L. (2020). The xylanase inhibitor TAXI-I increases plant resistance to Botrytis cinerea by Inhibiting the BcXyn11a xylanase necrotizing activity. Plants, 9.","DOI":"10.3390\/plants9050601"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/S0014-5793(98)00321-4","article-title":"Kunitz-type proteinase inhibitors from intact and Phytophthora-infected potato tubers","volume":"426","author":"Valueva","year":"1998","journal-title":"FEBS Lett."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1006\/bbrc.2001.5965","article-title":"A Bowman-Birk-type trypsin-chymotrypsin inhibitor from broad beans","volume":"289","author":"Ye","year":"2001","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"3129","DOI":"10.1016\/j.peptides.2006.07.013","article-title":"Isolation and characterization of a novel mung bean protease inhibitor with antipathogenic and anti-proliferative activities","volume":"27","author":"Wang","year":"2006","journal-title":"Peptides"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1726","DOI":"10.1016\/j.peptides.2006.01.020","article-title":"Psc-AFP, an antifungal protein with trypsin inhibitor activity from Psoralea corylifolia seeds","volume":"27","author":"Yang","year":"2006","journal-title":"Peptides"},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Vasconcelos, E.A., Santana, C.G., Godoy, C.V., Seixas, C.D., Silva, M.S., Moreira, L.R., Oliveira-Neto, O.B., Price, D., Fitches, E., and Filho, E.X. (2011). A new chitinase-like xylanase inhibitor protein (XIP) from coffee (Coffea arabica) affects Soybean Asian rust (Phakopsora pachyrhizi) spore germination. BMC Biotechnol., 11.","DOI":"10.1186\/1472-6750-11-14"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1094\/PHYTO-09-16-0346-R","article-title":"Peanut seed cultivars with contrasting resistance to Aspergillus parasiticus colonization display differential temporal response of protease inhibitors","volume":"107","author":"Bonacci","year":"2017","journal-title":"Phytopathology"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1471","DOI":"10.1104\/pp.106.4.1471","article-title":"Characterization and expression of an antifungal zeamatin-like protein (Zlp) gene from Zea mays","volume":"106","author":"Malehorn","year":"1994","journal-title":"Plant Physiol."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1111\/j.1745-4514.1995.tb00518.x","article-title":"Further characterization of the 12 kda protease\/alpha amylase inhibitor present in maize seeds","volume":"19","year":"1995","journal-title":"J. Food Biochem."},{"key":"ref_62","doi-asserted-by":"crossref","unstructured":"Apriyanto, A., Compart, J., and Fettke, J. (2022). A review of starch, a unique biopolymer\u2014Structure, metabolism and in planta modifications. Int. J. Exp. Plant Biol., 318.","DOI":"10.1016\/j.plantsci.2022.111223"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"60","DOI":"10.7897\/2230-8407.1206145","article-title":"A review on enzyme inhibitors","volume":"12","author":"Patadiya","year":"2021","journal-title":"Int. Res. J. Pharm."},{"key":"ref_64","first-page":"5","article-title":"Espectroscopia de infrarrojo con transformada de fourier (FT-IR) para an\u00e1lisis de muestras de harina de trigo, f\u00e9cula de ma\u00edz y almid\u00f3n de yuca","volume":"19","author":"Daza","year":"2021","journal-title":"Cienc. Tecnol. Aliment."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"105430","DOI":"10.1016\/j.foodhyd.2019.105430","article-title":"Preparation and characterization of cassava starch acetate with high substitution degree","volume":"100","author":"Cuenca","year":"2020","journal-title":"Food Hydrocoll."},{"key":"ref_66","doi-asserted-by":"crossref","unstructured":"Zi\u0119ba, T., Wilczak, A., Kobry\u0144, J., Musia\u0142, W., Kapelko-\u017beberska, M., Gryszkin, A., and Meisel, M. (2021). The annealing of acetylated potato starch with various substitution degrees. Molecules, 26.","DOI":"10.3390\/molecules26072096"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"923","DOI":"10.1016\/j.foodchem.2007.07.002","article-title":"Effect of acetylation on the properties of corn starch","volume":"106","author":"Chi","year":"2008","journal-title":"Food Chem."},{"key":"ref_68","doi-asserted-by":"crossref","unstructured":"Chi, C., Lian, S., Zou, Y., Chen, B., He, Y., Zheng, M., Zhao, Y., and Wang, H. (2023). Preparation, multi-scale structures, and functionalities of acetylated starch: An updated review. Int. J. Biol. Macromol., 249.","DOI":"10.1016\/j.ijbiomac.2023.126142"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"2100207","DOI":"10.1002\/star.202100207","article-title":"Antimicrobial starch-based film for food packaging application","volume":"74","author":"Romainor","year":"2022","journal-title":"Starch-St\u00e4rke"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1021\/ac60147a030","article-title":"Use of dinitrosalicylic acid reagent for determination of reducing sugar","volume":"31","author":"Miller","year":"1959","journal-title":"Anal. Chem."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"394","DOI":"10.3390\/analytica3040027","article-title":"Optimized spectrophotometry method for starch quantification","volume":"3","author":"Bahdanovich","year":"2022","journal-title":"Analytica"},{"key":"ref_72","doi-asserted-by":"crossref","unstructured":"Zhang, C., Du, M., Cao, T., and Xu, W. (2023). The effect of acetylation on the physicochemical properties of chickpea starch. Foods, 12.","DOI":"10.3390\/foods12132462"},{"key":"ref_73","unstructured":"Whistler, R.L. (1964). Acetylation. Methods in Carbohydrate Chemistry, Academic Press."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1002\/bmb.21522","article-title":"Enzyme kinetic parameters estimation: A tricky task?","volume":"49","author":"Aledo","year":"2021","journal-title":"Biochem. Mol. Biol. Educ."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/20\/9974\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T15:22:31Z","timestamp":1760973751000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/20\/9974"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,14]]},"references-count":74,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2025,10]]}},"alternative-id":["ijms26209974"],"URL":"https:\/\/doi.org\/10.3390\/ijms26209974","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,14]]}}}